Western blot analysis of extracts from various cell lines using CDK8 (P455) Antibody.
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Supplied in 10 mM sodium HEPES (pH 7.5), 150 mM NaCl, 100 µg/ml BSA and 50% glycerol. Store at –20°C. Do not aliquot the antibody.
For western blots, incubate membrane with diluted primary antibody in 5% w/v BSA, 1X TBS, 0.1% Tween® 20 at 4°C with gentle shaking, overnight.
NOTE: Please refer to primary antibody product webpage for recommended antibody dilution.
From sample preparation to detection, the reagents you need for your Western Blot are now in one convenient kit: #12957 Western Blotting Application Solutions Kit
NOTE: Prepare solutions with reverse osmosis deionized (RODI) or equivalent grade water.
Load 20 µl onto SDS-PAGE gel (10 cm x 10 cm).
NOTE: Volumes are for 10 cm x 10 cm (100 cm2) of membrane; for different sized membranes, adjust volumes accordingly.
* Avoid repeated exposure to skin.
posted June 2005
revised June 2020
Protocol Id: 10
CDK8 (P455) Antibody detects endogenous levels of total CDK8 protein.
Human, Mouse, Rat, Monkey
Polyclonal antibodies are produced by immunizing animals with a synthetic peptide corresponding to residues surrounding Pro455 of human CDK8. Antibodies are purified using protein A and peptide affinity chromatography.
The mammalian Mediator Complex is a multi-subunit protein complex that couples specific transcriptional regulators to RNA polymerase II (Pol II) and the basal transcription machinery. Interactions between distinct Mediator subunits and transcription factors allow for specific gene regulation (reviewed in 1).
Mediator complex interactions control various biological processes, including insulin signaling (2), NF-κB-dependent signaling (3), stem cell pluripotency and self renewal (4,5), and proliferation of colon cancer cells (6,7).
CDK8/Cyclin C, along with Med12 and Med13, constitute a subcomplex within the Mediator Complex thought to act as a molecular switch, inhibiting Pol II recruitment and transcription initiation (8,9). Expression of CDK8 abrogates E2F-1-dependent inhibition of β-catenin activity in colon cancer cells (9). High levels of CDK8 coincide with high β-catenin-dependent transcription in colon cancer cells, and their proliferation can be inhibited by suppressing CDK8 expression (8).
CDK8 can phosphorylate Ser727 on STAT1, which reduces natural killer (NK) cell toxicity (10,11). As such, inhibitors are being pursued as potential therapeutics to enhance NK cell activity and combat a variety of cancer types (12,13).
Explore pathways + proteins related to this product.
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